LOCUS       sequence-446510-        4950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low copy cloning vector with mCherry integrated in the multiple 
            cloning site (AmpR). There is a SmaI immediately upstream of mCherry
            start codon, which allows for C-terminal mCherry fusions..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4950)
  AUTHORS   Braun HG, Kanwal N, Rivera Lopez LF, Thomassin JL
  TITLE     Generation of a plasmid series for rapid sub-cloning and use in 
            various Enterobacteriaceae.
  JOURNAL   J Biosci Bioeng. 2024 Sep 6:S1389-1723(24)00253-6. doi: 
            10.1016/j.jbiosc.2024.08.006.
  PUBMED    39244484
REFERENCE   2  (bases 1 to 4950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biosci 
            Bioeng. 2024 Sep 6:S1389-1723(24)00253-6. doi: 
            10.1016/j.jbiosc.2024.08.006."
COMMENT     Sequence Label: pHBJT031
FEATURES             Location/Qualifiers
     source          1..4950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(95..886,887..955))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     718..737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(956..1060)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1128..1146
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1184..1206)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1306..1325
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1519..1541
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1533..1550
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1534..1550
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2342..2358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2342..2358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2355..2377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2366..2382
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2390..2396,2397..2414,2415..2420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2435..2456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2573..2590)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3476..4426)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      4474..4696
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg
       61 gcgggtctgc ggagtggtga atccgttagc gaggttacca atgcttaatc agtgaggcac
      121 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      181 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      241 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      301 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      361 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      421 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      481 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      541 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      601 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
      661 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
      721 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
      781 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
      841 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
      901 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
      961 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1021 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1081 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1141 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1201 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1261 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1321 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1381 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     1441 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     1501 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
     1561 tcggtaccct attatcatta cttgtacagt tcatccatac caccggtgct gtgacgaccc
     1621 tccgcgcgct catattgttc cacaatggtg tagtcttcgt tatggctggt gatgtctaac
     1681 ttaatattca cgttataggc acccggcaac tgaaccggtt ttttagcctt ataggtggtt
     1741 tttacttctg catcgtagtg gccaccgtcc ttcaacttca ggcgctgttt gatctcaccc
     1801 ttcagcgcac cgtcttccgg gtacatgcgc tcgctgctcg cttcccaacc catggtcttt
     1861 ttctgcatca ccgggccatc agacgggaag ttcgtaccac gcagcttaac tttatagatg
     1921 aactcgccat cttgcaggct gctgtcttgg gtaacggtca cgacaccgcc gtcctcgaaa
     1981 ttcataacac gctcccactt aaaaccctcc gggaaggaca acttcaggta gtccggaata
     2041 tctgccggat gtttaacgta cgctttgctg ccgtacataa actgcggaga caggatgtcc
     2101 caggcaaaag gcagtggtcc gcctttcgtg accttgagtt tagcagtttg cgtgccttca
     2161 tacggacggc cctcgccctc gccctcaatt tcgaactcgt gaccgttcac ggagccttcc
     2221 atgtgcactt taaaacgcat gaactctttg atgatcgcca tattatcttc ctctcctttt
     2281 gataccatcc cggggatcct ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat
     2341 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2401 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2461 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2521 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2581 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2641 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2701 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gttggagact tacccgaaat
     2761 ctgatttatt caacaaagcc actttcagta tggatttggg tgttacgggg gggggttggt
     2821 gcttaaaatc gcgctgagga tttctgatgg tgttaagcgg gcggttttga gatgtaaact
     2881 cgcccgttta acataatgga tcttgcgcgc accgcccgaa caccactcgc cacaaaaaac
     2941 cgccggaacg tccaaaagta cgggttttgc tgcccgcaaa cgggctgttc tggtgttgct
     3001 agtttgttat cagaatcgca gatccggctt cagccggttt gccggctgaa agcgctattt
     3061 cttccagaat tgccatgatt ttttccccac gggaggcgtc actggctccc gtgttgtcgg
     3121 cagctttgat tcgataagca gcatcgcctg tttcaggctg tctatgtgtg actgttgagc
     3181 tgtaacaagt tgtctcaggt gttcaatttc atgttctagt tgctttgttt tactggtttc
     3241 acctgttcta ttaggtgtta catgctgttc atctgttaca ttgtcgatct gttcatggtg
     3301 aacagctttg aatgcaccaa aaactcgtaa aagctctgat gtatctatct tttttacacc
     3361 gttttcatct gtgcatatgg acagttttcc ctttgatatg taacggtgaa cagttgttct
     3421 acttttgttt gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga
     3481 tccttccgta tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg
     3541 agaacgaacc attgagatca tacttacttt gcatgtcact caaaaatttt gcctcaaaac
     3601 tggtgagctg aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta
     3661 ggtaggaatc tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt
     3721 tctcaagttc ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat
     3781 cagtcgggcg gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt
     3841 tacttattgg tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa
     3901 gcattaacat gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct
     3961 tttgtgttag ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag
     4021 acttaacatg ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata
     4081 tctcttcact aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact
     4141 ggaaaatctc aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct
     4201 ctctggttgc tttagctaat acaccataag cattttccct actgatgttc atcatctgag
     4261 cgtattggtt ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg
     4321 ggttgagtag tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc
     4381 gactaatcgc tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt
     4441 ctaggtgatt ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc
     4501 ttttcctttg agttgtgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa
     4561 attctgctag accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc
     4621 aagtggttat aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca
     4681 gccctgtgta taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa
     4741 cgctgtttgc tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg
     4801 caagctcggg caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc
     4861 tgtctttttc gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa
     4921 tggcactaca ggcgcctttt atggattcat
//
